title: Characterization of PvuRts1I endonuclease as a tool to investigate genomic 5-hydroxymethylcytosine.
abstract: In mammalian genomes a sixth base, 5-hydroxymethylcytosine ((hm)C), is generated by enzymatic oxidation of 5-methylcytosine ((m)C). This discovery has raised fundamental questions about the functional relevance of (hm)C in mammalian genomes. Due to their very similar chemical structure, discrimination of the rare (hm)C against the far more abundant (m)C is technically challenging and to date no methods for direct sequencing of (hm)C have been reported. Here, we report on a purified recombinant endonuclease, PvuRts1I, which selectively cleaves (hm)C-containing sequences. We determined the consensus cleavage site of PvuRts1I as (hm)CN(11-12)/N(9-10)G and show first data on its potential to interrogate (hm)C patterns in mammalian genomes.
authors.affiliation: Ludwig Maximilians University Munich, Department of Biology and Center for Integrated Protein Science Munich, 82152 Planegg-Martinsried, Germany.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Szwagierczak
authors.forename: Andreas
authors.initials: A
authors.lastname: Brachmann
authors.forename: Christine S
authors.initials: CS
authors.lastname: Schmidt
authors.forename: Sebastian
authors.initials: S
authors.lastname: Bultmann
authors.forename: Heinrich
authors.initials: H
authors.lastname: Leonhardt
authors.forename: Fabio
authors.initials: F
authors.lastname: Spada
mesh_terms: D044503:5-Methylcytosine / Q000031:analogs & derivatives; D000818:Animals; D003596:Cytosine / Q000031:analogs & derivatives* / Q000032:analysis; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D053837:DNA Cleavage; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D016678:Genome; D051379:Mice
chemical_list: C011865:5-hydroxymethylcytosine; D044503:5-Methylcytosine; D003596:Cytosine; D004247:DNA; D004262:DNA Restriction Enzymes; C024369:endodeoxyribonuclease Rts1